Summary
The [H3]-labelled photo-activated analog of streptomycin (photo-Sm) is obtained as a result of the streptomycin reaction with 2-nitro, 4-azidobenzoylhydrazide and subsequent reduction with NaBH4 3. The analog retains the functional activity of the initial antibiotic as judged by two criteria: (1) it binds only to the 30S subparticle of ribosomes and (2) it inhibits the factor-free (“non-enzymatic”) PCMB-stimulated polyU-dependent system of translation (Gavrilova and Spirin, 1971).
After irradiation of the reaction mixture containing photo-Sm and either the 30S or 50S subparticles of ribosomes under similar conditions, the analog covalently binds chiefly to the 30S subparticle. Irradiation of the photo-Sm mixture with whole 70S ribosomes leads to a uniform distribution of a covalently bound label among the subparticles. A comparison of the effects obtained allows the conclusion that the analog is located on the interface of the ribosomal subparticles.
In the 30S subparticle the photo-Sm attacks mainly the protein component (more than 95% of all the covalently bound label). The proteins labelled by photo-reaction are identified as S7 (main), S14 (additional) and S16/S17 (minor).
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Communicated by H.G. Wittmann
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Girshovich, A.S., Bochkareva, E.S. & Ovchinnikov, Y.A. Identification of components of the streptomycin-binding center ofE. coli MRE 600 ribosomes by photo-affinity labelling. Molec. Gen. Genet. 144, 205–212 (1976). https://doi.org/10.1007/BF02428110
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DOI: https://doi.org/10.1007/BF02428110